Hepatogenic Differentiation of Human Induced Pluripotent Stem cells on Collagen-Coated Polyethersulfone Nanofibers

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Abstract

Many scientists have been fascinated with induced pluripotent stem cells (iPSCs) for cell replacement therapies. Nanofibrous biocompatible scaffolds have been shown to foster better cell adhesion and improve stem cell differentiation. In the current study, after fabrication using electrospinning technique and surface modifications, the characteristics of polyethersulfone (PES) nanofibers were determined by scanning electron microscopy, attenuated total reflection Fourier transform infrared spectroscopy, and 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide (MTT) assay. Then, the hepatogenic potential of iPSCs was evaluated using real-time reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry (ICC) after culture on collagen-coated polyethersulfone (PES/COL) scaffolds. After scaffolds characterization, analysis of two important definitive endoderm specific markers (Sox17 and Foxa2) using real-time RT-PCR and ICC indicated increase in their mRNA and protein levels after 5 days of hepatogenic induction. In addition, to determine hepatic differentiation of iPSCs cultured on PES/COL, the expression of albumin and α-fetoprotein was evaluated by ICC after 20 days. Real-time RT-PCR analysis showed increased expression of albumin, TAT, cytokeratin 19, and Cyp7A1 genes during the course of differentiation program. Finally, enzyme-linked immunosorbent assay analysis demonstrated an increased expression of albumin in the protein level after 28 days of differentiation. In conclusion, our results demonstrated that PES/COL nanofibrous scaffolds could be a proper substrate to significantly increase the hepatogenic differentiation potential of iPSCs and could also be introduced as a promising candidate for liver tissue engineering applications.

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